CD147 self-regulates matrix metalloproteinase-2 release in gingival fibroblasts after coculturing with U937 monocytic cells
CD147 self-regulates matrix metalloproteinase-2 release in gingival fibroblasts after coculturing with U937 monocytic cells
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DOI：
10.1002/jper.19-0278
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发表时间：
2019-10-21
影响因子：
4.3
通讯作者：
Fu, Earl
中科院分区：
文献类型：
--
作者：
Lai, Tat-Ming;Kuo, Po-Jan;Fu, Earl
背景 分化簇 147 (CD147) 是一种多功能糖蛋白，可作为成纤维细胞中基质金属蛋白酶 (MMP) 表达的诱导剂。最近在人牙龈成纤维细胞 (HGF) 和 U937 人单核细胞共培养中观察到 MMP-2 表达协同增强；然而，责任机制尚未完全建立。本研究的目的是评估与U937细胞共培养后HGF中可溶性CD147的释放及其对HGF中MMP-2表达增强的功能作用。方法采用酶联免疫吸附法测定培养基中CD147蛋白含量，实时聚合酶链反应检测HGFs和U937细胞中CD147和MMP-2 mRNA水平。通过酶谱法检测细胞释放的MMP-2的酶活性。选择Transwell共培养和条件培养基处理以排除HGF和U937细胞直接接触的影响。结果与U937细胞Transwell共培养并暴露于U937条件培养基后，HGFs中CD147蛋白和mRNA表达增强。通过共培养方法，HGF 中的 MMP-2 酶活性也显着增加。施用外源CD147增强了HGF中MMP-2的表达，而用抑制CD147表达的环孢菌素-A处理，降低了HGF中U937增强的MMP-2表达。结论 CD147可以与成纤维细胞相互作用，刺激与牙周细胞外基质降解相关的MMPs表达。这项研究表明，成纤维细胞释放的 CD147 可能在 HGF 中单核细胞增强的 MMP-2 表达中发挥作用。
Background Cluster of differentiation 147 (CD147) is a multifunctional glycoprotein that functions as an inducer of matrix metalloproteinase (MMP) expression in fibroblasts. Synergistically enhanced MMP-2 expression was recently observed in the coculture of human gingival fibroblasts (HGFs) and U937 human monocytic cells; however, the responsible mechanisms have not yet been fully established. The aim of this study was to evaluate the release of soluble CD147 in HGFs after coculturing with U937 cells and its functional effect on the enhancement of MMP-2 expression in HGFs. Methods Enzyme-linked immunosorbent assay was used to determine the amount of CD147 protein in media, whereas real-time polymerase chain reaction was performed to evaluate the mRNA levels of CD147 and MMP-2 in HGFs and U937 cells. The enzyme activities of MMP-2 released from cells were examined by zymography. Transwell coculturing and conditioned media treatments were selected to rule out the effect of direct contact of HGFs and U937 cells. Results The protein and mRNA expression of CD147 in HGFs were enhanced after transwell coculturing with U937 cells and exposure to U937-conditioned medium. MMP-2 enzyme activities in HGFs were also significantly increased by the coculturing methods. Administration of exogenous CD147 enhanced MMP-2 expression in HGFs, whereas treatment with cyclosporine-A, which inhibited CD147 expression, reduced U937-enhanced MMP-2 expression in HGFs. Conclusions CD147 can interact with fibroblasts to stimulate the expression of MMPs associated with periodontal extracellular matrix degradation. This study has demonstrated that CD147 released from fibroblasts might play a role in monocyte-enhanced MMP-2 expression in HGFs.